I utilized protocol from Willianne I.M. Vank et al.,2010 (J Biol Chem) without significant results.
I visualized other protocol, but different NBT (nitro-blue tetrazolium) and Riboflavin concentration, method and time exposure are reported. If someone can help me i would be grateful.
Daniil R. Petrenyov
Not me but my colleague did it.
Riboflavin under UV exposition will generate superoxide and SOD will cleaves it.
As I remember the all area of gel was blue but nor places of SOD activity localization. Since SOD is complex of two subunits (at least) non-denaturating conditions will be better.
Have you blue stain at all?
Usually NBT salts stable for decades, riboflavin may be not..
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